Anti-inflammatory Effect of Saikosaponin d on LPS-induced RAW264.7 Cell Inflammation
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Abstract:
A mouse model of lipopolysaccharide (LPS)-induced RAW264.7 cell inflammation was developed to investigate the in vitro anti-inflammatory mechanism of saikosaponin d (SSd). SSd demonstrated a significant inhibition of nitric oxide release from RAW264.7 cells at a concentration of 8 μmol/L (P<0.01) and suppressed mRNA level expression of inflammation-related genes, including inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), interleukin-6, and tumor necrosis factor-α, in a dose-dependent manner at concentrations of 4 and 8 μmol/L (P<0.01). Western blot analysis revealed that SSd effectively reduced the protein level expression of iNOS and COX-2. SEM images confirmed that SSd alleviated LPS-induced morphological changes in RAW264.7 cells. Additionally, SSd inhibited the expression and phosphorylation levels of key proteins involved in inflammation-related signaling pathways, including toll-like receptor 4 (TLR4), IκB kinase, nuclear factor κB (NF-κB), and glycogen synthase kinase-3β (GSK3β). These findings indicate that SSd strongly inhibits LPS-induced RAW264.7 cell inflammation by targeting the TLR4/NF-κB and GSK3β signaling pathways. These findings contribute to our enhanced understanding of the anti-inflammatory effects of SSd.
